1. Julian Avila, Timothy Devarenne Characterization of the interaction of the negative regulator of cell death Adi3 with the SnRK complex

2. Pseudomonas syringae Photo: Gordon Vrdoljak, Electron Microscopy Laboratory, U.C. Berkeley Bacterial speck disease DISEASE PROGRAMMED CELL DEATH (PCD) ?

3. AvrPto-dependent Pto-interacting protein 3 Adi3 Devarenne et al. (2006) EMBO J. Adi3 is a protein kinase Silencing of Adi3 causes localized cell death Kinase defficient (K337Q) Constitutively active (S539D)

4. How is Adi3 a cell death suppressor? Bait Adi3 Prey PDK1 Bicoid Atg8 At3g13230 Thioredoxin E3 Ubiquitin Ligase SnRK autophagy putative TF cellular reduction /oxidation state ubiquitination Kinase Y2H screen using Adi3 as a bait SnRK SnrK1 pJG4-5 pEG202 SnRK Adi3 SnRK Prey Bait (PULLED DOWN) Kinase defficient (K337Q) Constitutively active (S539D)

5. SNF (Sucrose Non Fermenting) SNF4 SIP1 GAL83 IT IS THE CATALYTIC SUBUNIT OF THE SNF1 RELATED KINASE COMPLEX (β)(β) (γ)(γ) What is SnRK? (Sucrose non-fermenting related kinase) (α)(α) SnRK AMPK (AMP Dependent-Kinase) AKIN SnRK

6. SnRK > GAL83 > SnRK > GAL83 > Adi3 > Is SnRK an active kinase? ATP ADP Is Adi3 phosphorylated by the complex? P P Does Adi3 phosphorylate GAL83 (β)? (γ)(γ) (β)(β) ? (α)(α) (β)(β) (γ)(γ) ATP ADP GAL83 > Adi3 > GAL83 > Adi3 > SnRK

7. α-MBP Does Adi3 phosphorylate both GAL83 (β1) and SIP1 (β2) ? Is Adi3 able to interact with both β subunits? GAL83/SIP1 > Adi3 > GAL83/SIP1 > Adi3 > P (β)(β) α-GST ? ATP ADP SIP1 GAL83

8. P? Which residue(s) is(are) phosphorylated on GAL83 by Adi3? - Mutagenesis screen on serines.

9. S26 ? SnRK-complex mediated stress responses? Programmed cell death? SnRK-complex mediated stress responses? Programmed cell death? ?

10. Andrey Belyanin Devarenne Lab Members: Anna Nelson Joel Gray Taylor Weiss Julissa Ek-Ramos Remington Fenter Committee Members: Dr. Tim Devarenne Dr. J.P. Pellois Dr. Paul Straight Dr. Michael Kolomiets Funding: Acknowledgements